ISSN 2756-3707
International Journal of Histology and Cytology ISSN 2447-9535 Vol. 6 (1), pp. 001-005, January, 2019. © International Scholars Journals
Full Length Research Paper
Expression of complete rhoptry protein 2 (ROP2) gene of Toxoplasma gondii in eukaryotic cell
K. Hosseinian Khosroshahi1, F. Ghaffarifar1*, Z. Sharifi2 and A. Dalimi1
1Department of Parasitology, Medical Sciences Faculty, University of Tarbiat Modarres, Tehran, Iran.
2Research Center of Iranian Blood Transfusion Organization, Tehran, Iran.
Accepted 16 November, 2018
Abstract
Toxoplasma gondii is the intracellular protozoan parasite responsible for animal and human toxoplasmosis. In immunodeficient patients, chronic infection with T. gondii can reactivate and produce encephalitis, which is often lethal. ROP2 (rhoptry protein of T. gondii) is one of the most important interferer in organelle and PVM blending. ROP2 protein is recognized by clone T-cell (Tcc32) in human body and also has epitope for B-cell. All of these characteristics of ROP2 makes it a candidate for cocktail vaccine and recombinant vaccine against toxoplasmosis. We described the expression of the gene which encodes the complete rhoptry protein 2 (ROP2) of T. gondii in CHO cells and confirmed it by SDS-PAGE and Western blot analysis. In the present work, genomic DNA of T. gondii was extracted and used for amplifying of ROP2 gene as a template. Then PCR product was cloned into pTZ57R/T vector, and plasmid containing ROP2 gene (pT-ROP2) was extracted from transformed bacteria and sequenced. We hope to use from this recombinant plasmid (pT-ROP2) to make DNA vaccine against toxoplasmosis.
Key words: Cloning, sequencing, Toxoplasma gondii, ROP2.